Journal of AOAC International最新文献

Background: The 2018 Farm Bill defines hemp as Cannabis sativa L. with a tetrahydrocannabinol (THC) concentration not more than 0.3% on a dry weight basis where THC is implied to be total THC (THCTotal) including both acid (Δ9-THCA) and neutral (Δ9-THC) forms.

Objective: Effect of temperature and mass were studied to determine the quickest time to prepare fresh hemp suitable for grinding without affecting THCTotal. Proficiency testing program data were also analyzed to compare THCTotal contents on a dry versus wet weight basis.

Methods: Fresh hemp of 150 and 400 g were freeze-dried or heat-treated at 50 °C, 60 °C, and 70 °C for various lengths of time up to 72 hours. Ground hemp was analyzed for moisture content via AOAC 934.01 and THCTotal via liquid chromatography/mass spectrometer (LC/MS) or gas chromatography with flame ionization detection (GC/FID). A data set analyzed from a proficiency testing program included moisture and THCTotal from 20 to 67 laboratories on 12 analytical samples circulated from 2020 through 2022.

Results: The quickest drying time of 24 h occurred at 70 °C with 150 g. These conditions did not affect THCTotal content on a dry weight basis. Twelve proficiency testing program analytical samples ranged from 5.8 to 11.4% moisture and 0.144 to 0.399% THCTotal on a wet weight basis. An increase in reported THCTotal on a dry weight basis compared to a wet weight basis ranged from 0.002 to 0.027% with a significant difference occurring in only one analytical sample.

Conclusions: Suitable condition for drying hemp without altering THCTotal content was 24 h at 70 °C. Determining residual moisture in oven-dried ground hemp to calculate THCTotal content on a dry weight basis provides no benefit given minimal difference between contents on dry versus wet weight bases, interlaboratory variability, and terpene volatilization.

Background: Acrylamide (AA) is a process contaminant naturally formed during the cooking of starchy food at high temperatures. Considering existing risks of misquantification inherent to the analysis of AA, an AOAC initiative raised the need for a consensus standard to determine AA in a broad variety of food.

Objective: A quantitative LC-MS/MS method for AA determination in food was validated in a single-laboratory study. Targeted performance requirements in terms of target matrixes, limit of quantification, recovery, and precision were as defined per Standard Method Performance Requirement (SMPR®) 2022.006.

Method: The proposed method derives from EN 16618:2015 standard pending modifications brought to the (1) sample preparation (simplified, potentially automated); (2) scope of application (significantly extended); and (3) LC conditions (improved selectivity). Confirmatory detection of AA is conducted by LC-MS/MS in the Selected Reaction Monitoring mode (SRM), and isotopic dilution was applied for quantification approach using either 2,3,3-d3-acrylamide (d3-AA), or 13C3-2,3,3-d3-acrylamide (13C3-d3-AA) as labeled internal standard.

Results: A total of 16 laboratory samples from nine matrix categories were included in the validation process. A full validation was conducted on coffee (instant, roast), infant cereal, cocoa powder, pet food (croquettes), tea (green tea), spices (black pepper), and nuts (roasted almonds) with satisfactory performances both in terms of recovery (97-108%) and precision (RSDr and RSDiR <12%). The method applicability was further demonstrated through the analysis of quality control materials and reference materials including French fries, potato crisps, vegetable crisps, instant coffee, infant food, and biscuits (cookies), with accuracy values determined within a 94-107% range.

Conclusions: The performances of the presented method are in agreement with the acceptance criteria stipulated in SMPR 2022.006.

Highlights: The Expert Review Panel for acrylamide approved the present method as AOAC Official First Action 2023.01.

Background: Analytical quality by design (AQbD) affords a systematic scaffolding to triumph a continuously validated, robust assay as well as life cycle management. The resuscitative repurposed drug favipiravir, an oral drug approved for reemerging pandemic influenza in Japan in 2014, is used for the treatment of life-threatening pathogens such as Ebola, Lassa virus, and currently COVID-19. Favipiravir is gaining a great deal of medical importance due to its pharmaceutical applications.

Objective: To develop and validate a risk-based stability-indicating RP-HPLC method employing an AQbD approach using Central Composite Design (Design Expert Software 13.0) for the estimation of favipiravir.

Method: The Quality Target Product Profile optimized were flow rate and mobile phase composition, thus assessing the critical analytical attributes (retention time, tailing factor, and number of theoretical plates) as the constraints of method robustness. The proposed technique was optimized with a C18 (150 × 4.6 mm, 5 µm) column and 0.1% orthophosphoric acid buffer-acetonitrile (50:50, v/v) as the mobile phase at a flow rate of 1 mL/min using diode-array detector (230 nm) eluted favipiravir at 2.3 min.

Results: The optimized method validated as per ICH guideline Q2 (R1) was found to be eco-friendly, simple, precise (RSD 0.0051-1.2%), accurate (99.86-100.22%), linear (25-150 µg/mL), rugged (RSD 0.70%), and robust (RSD 0.6-1.6%) with a limit of detection and limit of quantitation of 1.140 µg/mL and 4.424 µg/mL, respectively.

Conclusion: Forced degradation studies (acidic, alkaline, thermal, photolytic, and oxidative conditions) revealed the suitability of the AQbD method for the analysis of favipiravir in tablet formulation.The developed and validated AQbD method is less time consuming and can be used in the industry for routine quality control/analysis of bulk drug and marketed Favipiravir products.

Highlights: A robust Design of Experiment enhanced stability-indicating analytical method was developed and validated for the estimation of favipiravir. Furthermore, the contemporary method would aid in extending the analysis of favipiravir in other formulations.

Background: The Qianlie Shule capsule is a classical Chinese medicine compound preparation frequently used in therapeutic settings to alleviate astringent pain in the urethra, prostatic hypertrophy, and chronic prostatitis or urinary frequency. However, a comprehensive analysis of the chemical composition of Qianlie Shule capsules has not been reported.

Objective: To establish a quick and effective analytical method based on hybrid quadrupole-orbitrap mass spectrometry ultrahigh-performance liquid chromatography (UPLC-Q-Orbitrap-MS/MS) for the identification and characterization of chemical components in Qianlie Shule capsules.

Method: Using ultrahigh-performance liquid chromatography with hybrid quadrupole-orbitrap mass spectrometry and data post-processing, the samples of Qianlie Shule capsules were examined. First, the whole extract of the Qianlie Shule capsules was separated using a UPLC machine, and the fragmentation data were collected in both positive and negative ion mode. The target molecule is then quickly identified by comparing the fragmentation information of the neutral loss (NLs) and characteristic fragments (CFs) reported in the literature.

Results: A total of 145 chemical components were identified. It includes flavonoids, triterpenoids, phenylpropanoids, organic acids, alkaloids, phenylethanoids, iridoids, and anthraquinones.

Conclusions: This study is a method for the rapid qualitative analysis of the chemical composition of Qianlie Shule capsules, which provides a method for the rapid, sensitive, and high-throughput identification of the prescription components of Chinese medicine.

Highlights: Systematic identification of the chemical composition of QLSL capsules provides a theoretical basis for studying the substance basis of QLSL capsules and the improvement of the quality control level.

While current analytical methodologies can readily identify cannabis use, definitively establishing recent use within the impairment window has proven to be far more complex, requiring a new approach. Recent studies have shown no direct relationship between impairment and Δ9-tetra-hydrocannabinol (Δ9-THC) concentrations in blood or saliva, making legal "per se" Δ9-THC limits scientifically unjustified. Current methods that focus on Δ9-THC and/or metabolite concentrations in blood, saliva, urine, or exhaled breath can lead to false-positive results for recent use due to the persistence of Δ9-THC well outside of the typical 3-4 h window of potential impairment following cannabis inhalation. There is also the issue of impairment due to other intoxicating substances-just because a subject exhibits signs of impairment and cannabis use is detected does not rule out the involvement of other drugs. Compounding the matter is the increasing popularity of hemp-derived cannabidiol (CBD) products following passage of the 2018 Farm Bill, which legalized industrial hemp in the United States. Many of these products contain varying levels of Δ9-THC, which can lead to false-positive tests for cannabis use. Furthermore, hemp-derived CBD is used to synthesize Δ8-THC, which possesses psychoactive properties similar to Δ9-THC and is surrounded by legal controversy. For accuracy, analytical methods must be able to distinguish the various THC isomers, which have identical masses and exhibit immunological cross-reactivity. A new testing approach has been developed based on exhaled breath and blood sampling that incorporates kinetic changes and the presence of key cannabinoids to detect recent cannabis use within the impairment window without the false-positive results seen with other methods. The complexity of determining recent cannabis use that may lead to impairment demands such a comprehensive method so that irresponsible users can be accurately detected without falsely accusing responsible users who may unjustly suffer harsh, life-changing consequences.

Background: The presentation of rhinitis has drawn increasing attention in recent years due to the possibility of overlap or confusion between allergic rhinitis symptoms and those of COVID-19. Azelastine hydrochloride (AZH) and mometasone furoate (MOF) are two of the most efficient combinations for enhancing the symptoms of seasonal allergic rhinitis.

Objective: This work concerns applying and validating different accurate and simple spectrophotometric approaches for simultaneous quantification of the binary mixture of AZH and MOF in raw material, laboratory-prepared mixtures, and pharmaceutical preparation. Moreover, assessment of the environmental impact of the applied approaches on the environment was also a key goal of this study.

Methods: AZH was determined using the direct spectrophotometric (D0) method, while four reliable spectrophotometric approaches namely, induced dual wavelength (IDW), ratio subtraction (RS), ratio difference (RD), and ratio derivative (1DD) were used for MOF determination.

Results: The methods were validated in line with the International Conference of Harmonization standards. In the AZH range of (5-56 µg/mL) and MOF range of (2-20 µg/mL), the linearity of the proposed approaches was investigated with high accuracy findings. There were no significant differences between the obtained results and those of the reported method when compared statistically. Furthermore, the applied spectrophotometric methods were deemed to be eco-friendly according to Green Analytical Procedure Index (GAPI) and Analytical Greenness Calculator (AGREE) assessment metrics.

Conclusions: The applied spectrophotometric methods are simpler, more eco-friendly, and take a shorter time to precisely estimate many measurements compared to the only reported chromatographic analysis.

Highlights: Neither publications of novel spectrophotometric methods nor reported green ones have been available for simultaneous determination of the binary mixture of AZH and MOF, so this work has a great significance and novelty in the area of pharmaceutical analysis.

Background: Ponceau 4R (E124) and carmoisine (CMS; E122) are frequently utilized azo synthetic dyes in the food industry owing to their aesthetically pleasing coloration and broad consumer acceptability. It is imperative to prioritize environmentally favorable technologies for quantifying these dyes, as excessive consumption of these poses significant health risks.

Objective: The primary objective of this research was to establish a reversed-phase (RP)-HPLC method that could simultaneously detect Ponceau 4R and CMS, implementing green analytical chemistry (GAC) and analytical quality by design (AQbD), using an ultrasound-assisted extraction (UAE) technique in commercial food samples.

Methods: An Agilent Eclipse Plus column (C18, 250 × 4.6 mm id, 5 µm) was utilized for effective separation with a mobile phase of ethanol-acetate buffer pH 5 (60:40, v/v), flow rate of 1 mL/min, and detection wavelength of 515 nm. Critical variables selected for method optimization were ethanol percentage and flow rate, determined using central composite design (CCD). In order to adhere to the 12 principles of green chemistry, hazardous solvents were substituted with ethanol, which is distinguished by its ease of use, effectiveness, and ecological sustainability. The greenness assessment was conducted utilizing the green analytical procedure index (GAPI), analytical eco-scale (AES), and analytical greenness metrics (AGREE).

Results: The respective retention times for Ponceau 4R and CMS were 2.276 and 3.450 min. The recovery rate of Ponceau 4R and CMS fluctuated between 70% and 102% and 80% and 102%, respectively, across various marketed food samples. The procedure passed validation in accordance with the International Conference on Harmonization Q14 guidelines.

Conclusion: The devised method demonstrates that the validation parameters like linearity, precision, sensitivity, and reproducibility are within the specified limits of ICH guidelines. The greenness assesment tools GAPI, AES, and AGREE produced the most favorable results.

Highlights: In future, environmentally sustainable, solvent-based, robust AQbD methodologies for assessing varieties of food colorants may be adopted and improved commercially.

Background: Food allergen cross-contact during food preparation and production is one of the causes of unintentional allergen presence in packaged foods. However, little is known about allergen cross-contact in shared frying or roasting oil, which prevents the establishment of effective allergen controls and may put allergic individuals at risk. To better understand the quantity of allergen transferred to frying oil and subsequent products, an analytical method is needed for quantifying protein in oil that has been exposed to frying/roasting conditions.

Objective: The goal of this study was to develop a parallel reaction monitoring LC-MS/MS method to quantify the amount of cashew protein in shared roasting oil.

Methods: The sample preparation method was evaluated to improve protein extractability and peptide performance. Four quantitative peptides representing cashew 2S and 11S proteins were selected as targets based on their sensitivity, heat stability, and specificity. A calibration strategy was developed to quantify the amount of total cashew protein in oil. Method performance was evaluated using a heated cashew-in-oil model system.

Results: The method showed high recovery in oil samples spiked with 100 or 10 parts per million (ppm) total cashew protein heated at 138 or 166°C for 2-30 min. Samples (100 ppm total cashew protein) heated for 30 min had more than 90% recovery when treated at 138°C and more than 50% when heated at 166°C.

Conclusion: The method is fit-for-purpose for the analysis of cashew allergen cross-contact in oil.

Highlights: A novel MS-based method was developed that can accurately quantify the amount of cashew protein present in heated oil.

Background: Emergence and dissemination of antibiotic resistance is one of the major risks associated with the rampant usage of antibiotics in food-producing animals including aquaculture.

Objective: To determine Epidemiological Cut-OFF (ECOFF) values of heterotrophic bacterial populations from shrimp culture environments against five different antibiotics.

Methods: In this present study, bacterial samples were isolated from Penaeus vannamei culture environment in different locations of Andhra Pradesh, which is the aquaculture hub of India. The bacterial isolates were assessed for antibiotic resistance towards five antibiotics belonging to different classes (oxytetracycline, chloramphenicol, erythromycin, ciprofloxacin, and co-trimoxazole) by the disc diffusion method. Determination of Epidemiological Cut-OFF (ECOFF) values and analysis by employing normalized resistance interpretation (NRI) was carried out.

Results: The most dominant bacterial populations from shrimp culture were Vibrio spp. (pathogenic bacteria) followed by Bacillus spp. (probiotic bacteria). The bacterial isolates showed highest resistance towards oxytetracycline (overall 23.38%) and in location L6 (59.4%) followed by co-trimoxazole (31.1%). ECOFF values calculated by employing NRI showed that the disc diffusion data were distributed in a normalized manner. The maximum ECOFF value was obtained for ciprofloxacin (23.32 mm), while the minimum value was observed for oxytetracycline (9.05 mm). The antibiotic resistant phenotypes showed that the majority of the heterotrophic bacterial isolates (>60%) belonged to the non-wild type phenotype and primarily towards oxytetracycline (90%).

Conclusion: The presence of non-wild antibiotic-resistant phenotypes of heterotrophic bacterial populations (which include not only pathogenic bacteria but also probiotic bacteria) indicates that shrimp culture ponds may be a reservoir for drug-resistant bacteria and there is a greater risk associated with transmission of resistant genes across bacterial flora.

Highlights: NRI analysis of antibiotic disc diffusion data of heterotrophic bacterial populations in shrimp aquaculture environments revealed that majority of them belonged to non-wild type (90%) paticularly to oxytetracycline in comparison to other studied antibiotics (chloramphenicol, erythromycin, ciprofloxacin and co-trimoxazole).

Background: Myrcene and cymene, aromatic monoterpenes found in plants and essential oils, possess distinctive aromatic qualities. However, their volatility and limited solubility pose challenges in precise handling and formulation. Meanwhile, nanoemulsions emerge as promising drug delivery systems, improving the bioavailability and stability of these active ingredients.

Objective: This article aimed to develop an HPLC method for the quantification of two monoterpenoids, p-cymene and myrcene, in nanoemulsions.

Method: The method used a Phenomenex® Synergi™ Fusion-RP column (150 mm × 4.6 mm id, 4 μm particle size) on an HPLC system with isocratic elution. The mobile phase was composed of acetonitrile and water (60:40, v/v) and was validated in terms of specificity, linearity, accuracy, precision, robustness, and selectivity.

Results: The method provided accurate and precise results with a correlation coefficient of 0.999 and RSD values of less than 2%. The method can be used for quality control of nanoemulsions containing these monoterpenoids and as a reference for future studies on their efficacy and stability.

Conclusions: The study demonstrates the feasibility of using HPLC for the quantification of monoterpenoids in nanoemulsions and its potential as a quality control tool for nanoemulsion-based drug delivery systems.

Highlights: The method's accuracy, precision, and reliability, as evidenced by high correlation coefficients and low RSD values, underscore its suitability for ensuring the consistent formulation of these monoterpenoid-containing nanoemulsions, while also serving as a reference point for future research endeavors in this field.